DNA extraction from dried tissue - (Apr/17/2008 )

Hi, i have to extract DNA from dried and old Tissue (1930´s/1960`s). It was material we got from a museum. So far i have tried the Qiagen DNeasy tissue Kit and ArchivePure DNATissue Kit from 5 Prime.It didn´t work at all. Because it´s really difficult to get fresh tissue or blood from these animals i have to give it another try. Has anyone a suggestion what else i could use ?

Thanks in advance , cissy

-cissi-

QUOTE (cissi @ Apr 17 2008, 12:17 PM)

Hi, i have to extract DNA from dried and old Tissue (1930´s/1960`s). It was material we got from a museum. So far i have tried the Qiagen DNeasy tissue Kit and ArchivePure DNATissue Kit from 5 Prime.It didn´t work at all. Because it´s really difficult to get fresh tissue or blood from these animals i have to give it another try. Has anyone a suggestion what else i could use ?

Thanks in advance , cissy

There are various protocols for DNA extraction from museums species. But the actual conditions of the material varies, the amounts (yield) can be very different, DNA can from a bit to completely degraded. One critical issue is the method of conservation, if species were dried, this can work well (in small species) as drying is fast, in larger species it may not work or only partly (e .g in the legs). Alcohol can work (though some ingredients of it can be fatal, it can be diluted by body fluids). Formalin I have no experience. Since all DNA is more or less degraded or at least cut, frequently occurring DNA and shorter DNA fragments are better to use (i.e. mDNA or smaller markers). One last point: How did you checked if your protocol worked? If only a gel there might be not enough DNA to see it, nevertheless PCR will work. And have a look for PCR inhibitors, an additional purification step might be useful. Papers:ANCIENT DNA STUDIES IN PHYSICAL ANTHROPOLOGY - Dennis H. O’Rourke et al. - Annu. Rev. Anthropol. 2000. 29:217–42.Comparison and optimization of ancient DNA extraction - Nadin Rohland & Michael Hofreiter - BioTechniques. 2007. 42:343-352